As an essential step in mammalian gene expression, pre-mRNA splicing is closely regulated by many cis-elements and trans-acting splicing factors. It was estimated that at least 15% of point mutations that result in human genetic diseases disrupted splicing. The proposed research investigates the function of disease-associated genetic variants from the perspectives of pre- mRNA splicing and gene expression pathways. We will use a synergistic combination of computational methods with molecular genetic and functional genomic approaches. We will focus on two specific aims: (1) to develop a computational scheme for the identification of genetic variants that can potentially alter splicing, followed by experimental validation of allele- specific abnormal splicing using minigene reporters and cultured cell lines;(2) to identify genetic interaction networks and pathways specific to alternatively spliced isoforms induced by genetic variants by combining isoform-specific experimental perturbation and known interaction networks. These studies will allow an improved understanding of the consequences of genetic variants in individual gene expression and genetic interaction networks. PUBLIC HEALTH RELEVANCE: Narrative Alternation of normal splicing can significantly affect gene expression and contribute to human diseases. The proposed research aims to identify genetic mutations and polymorphisms that induce abnormal splicing and to investigate how such splicing abnormality is involved in genetic interaction networks. A better understanding in the above aspects will shed light on functional mechanisms underlying complex human diseases, such as diabetes, lupus and various types of cancer.